The goal of this proposal is to purchase/acquire and implement a Two-photon microscope which will primarily be utilized to strengthen the neuroscience research at Wright State, and permit key investigators to perform studies they would otherwise be unable to accomplish. We have assembled a team of highly accomplished researchers who will constitute the major users of the requested instrument and all of which are PIs on NIH funded grants (R01s). Dr. Francisco Alvarez will use the requested instrument to study Generation and functional role of Renshaw cells in the developing spinal cord. This study has broad implications on motor development and reconfiguration and recovery of function of neurons in adults after spinal cord injuries that disrupt these intraspinal circuits. Dr. Francisco Alvarez-Leefman will utilize the microscope to understand the cellular and molecular mechanisms underlying cell volume control in neurons and glial cells. Dr. Robert Fyffe will utilize the microscope to investigate the dynamics of surface membrane ion channel localization following altered activity or axonal injury, a project that also has implications in the field of spinal cord/nerve injury and recovery. Dr. Robert Putnam will use the requested instrument to study the role of Intracellular Ca and pH in chemosensitive signaling in dendrites of brainstem neurons. A greater understanding of central chemosensitive signaling will open new targets for treatments aimed at altering respiratory drive in such disorders as central hypoventilation syndrome, SIDS, and sleep apnea. The instrument will be housed and fully supported in a core facility in the recently created Comprehensive Neuroscience Center, where, in addition to these four major users, there will be opportunity for the instrument to be used by additional staff/Minor users/ Junior Investigators for cellular and molecular studies. Based on our current expertise as well as our collaboration with Dr Michael O'Donovan at NIH (who has invited Dr. Alvarez for a month to use a two-photon microscope at NIH) we believe that we will master the use of the Two-photon microscope and will be able to make the most of its potential. A Two-photon microscope will enhance the Imaging Core Facility at Wright State which currently houses two confocal microscopes and a transmission electron microscope, all networked to off-line image analysis workstations. The technical expertise to be gained will be utilized to further advance other research endeavors at WSU. PUBLIC HEALTH RELEVANCE: A Two-Photon Microscope will enhance the Neuroscience core at Wright State University and promote state- of-the art scientific research. The research proposed here will make use of this instrument to its full capacity, leading to an understanding of both neurophysiologic and neuropathologic mechanisms of high relevance to nervous system function in health and disease or injury conditions.